International Biodeterioration & Biodegradation xxx (2015) 1e9

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International Biodeterioration & Biodegradation

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Biological nutrient removal by recirculating aquaponic system:

Optimization of the dimension ratio between the hydroponic &
rearing tank components
Su Shiung Lam a, c, *, Nyuk Ling Ma b, Ahmad Jusoh a, Mohd Azmi Ambak c
a
Eastern Corridor Renewable Energy Group (ECRE), School of Ocean Engineering, University Malaysia Terengganu, 21030 Kuala Terengganu, Terengganu,
Malaysia
b
School of Fundamental Science, University Malaysia Terengganu, 21030 Kuala Terengganu, Terengganu, Malaysia
c
Institute of Tropical Aquaculture, University Malaysia Terengganu, 21030 Kuala Terengganu, Terengganu, Malaysia

a r t i c l e i n f o a b s t r a c t

Article history: Marble goby (Oxyeleotris marmorata Bleeker) was cultured in a recirculating aquaponic system (RAS)
Received 30 December 2014 containing a hydroponic tank grown with water spinach (Ipomoea aquatica). The influence of component
Received in revised form ratio (hydroponic tank volume to rearing tank volume) on the fish growth, vegetable yield, and nutrient
8 March 2015
removal was investigated. Increased fish growth (2.4 g/day), vegetable yield (22 kg/harvest), and nutrient
Accepted 9 March 2015
Available online xxx
removal (83% ammonia-N removal, 87% nitrite-N removal, 70% nitrate-N removal, 60% removal of total
phosphorus, 88% removal of total suspended solid, 63% removal of 5-day biochemical oxygen demand)
were observed at high component ratio (3 m3/m3). Component ratio was found to have a significant
Keywords:
Nutrient removal
influence on nutrient removal and production of marble goby and water spinach in RAS. A component
Aquaponic ratio of
3 m3 of hydroponic tank volume to 1 m3 of fish rearing tank volume showed advantages in
Hydroponic improving the production of the fish and vegetable and removing the nutrient wastes, TSS, and BOD5
Aquaculture generated from the culture of the fish. The results indicate that RAS show exceptional promise as a means
Marble goby to the reduction of biological nutrients accumulated in aquaculture wastewater and in turn providing a
good water quality environment for fish culture.
© 2015 Elsevier Ltd. All rights reserved.

Introduction
Marble goby (Oxyeleotris marmorata Bleeker) is a freshwater
food fish that commands a high price in Southeast Asia, ranging
from a wholesale price of 20e30 US dollars/kg (Lam et al., 2008;
Chew et al., 2009; Loo et al., 2013). The fish is conventionally
cultured in lakes, rivers, and ponds in countries such as Malaysia,
Thailand, and Vietnam (Luong et al., 2005; Chew et al., 2009; Loo
et al., 2013). Some attempts to culture the fish in earthen ponds
and cages have failed because of the disease problem caused by
Aeromonas hydrophila and Lernaea cyprinacea (anchor worm), the
slow growth of the fish during juvenile stage, and the high fish
mortality rate (Ang Kok, 1980; Cheah et al., 1994; Tng et al., 2008;
* Corresponding author. Eastern Corridor Renewable Energy Group (ECRE),
School of Ocean Engineering, University Malaysia Terengganu, 21030 Kuala
Terengganu, Terengganu, Malaysia. Tel.: þ60 (9) 668 3844; fax: þ60 (9) 668 3991.
E-mail addresses: lam@umt.edu.my (S.S. Lam), nyukling@umt.edu.my (N.L. Ma),
ahmadj@umt.edu.my (A. Jusoh), ambak@umt.edu.my (M.A. Ambak).
Nhi et al., 2010; Idris and Amba, 2011). It is thought that these
problems are derived from the poor water quality and the lack of
appropriate control present in the conventional culture of marble
goby. Therefore, it is important to find an alternative culture
technique that could rectify these deficiencies in order to ensure
better production of the fish.
Recirculating aquaponic system (RAS), a water recirculating
system that is designed to produce both fish and plants, can
potentially be a good culture technique for the fish as it ensures a
good controlled culture condition by providing better control of
water quality, reduced water usage, improved waste management
and nutrient recycling (Hamlin et al., 2008; Endut et al., 2009;
Martins et al., 2010; Lam et al., 2014). In RAS, the biological
nutrient wastes excreted by fish (e.g. ammonia) and those gener-
ated from the microbial breakdown of fish feed (nitrite, nitrate) are
absorbed by plants as nutrients for growth, and thus this method
allows the removal of undesirable nutrient wastes from the water
by plants and the water can then be reused for fish culture. These
could potentially lead to faster growth and higher production of

http://dx.doi.org/10.1016/j.ibiod.2015.03.012
0964-8305/© 2015 Elsevier Ltd. All rights reserved.

Please cite this article in press as: Lam, S.S., et al., Biological nutrient removal by recirculating aquaponic system: Optimization of the dimension
ratio between the hydroponic & rearing tank components, International Biodeterioration & Biodegradation (2015), http://dx.doi.org/10.1016/
j.ibiod.2015.03.012
2 S.S. Lam et al. / International Biodeterioration & Biodegradation xxx (2015) 1e9
both the fish and plants. The use of RAS techniques has been re-
ported to be highly efficient as it utilizes the generated fish waste as
nutrients for the plants and thus providing a symbiotic environ-
ment for producing fish and plants in a closed system (Martins
et al., 2010). However, the need for viable methods for protection
of plants grown in RAS is currently under development as there has
not been a fish-safe insecticide or fungicide developed for use in
aquaponics (Pilinszky et al., 2015). There is also a need to design the
RAS to maintain the balance of nutrient production and uptake in
order to ensure effective nutrient removal (Buzby and Lin, 2014).
Owing to the tremendous market demand for marble goby and
the limitations and poor fish yield shown by conventional culture
methods, it was thought useful to investigate the development of a
RAS for efficient control of water quality and improved fish pro-
duction from the culture of the fish. The distinct advantages shown
by RAS may overcome the poor growth and disease problem shown
by conventional cage and pond culture systems and lead to the
potential for the greater production of the fish. So far, limited in-
formation is available on the characteristics of the culture of marble
goby. Studies on the application of RAS have been reported in the
culture of other species, such as seabass (Franco-Nava et al., 2004),
African catfish (Endut et al., 2010) and carp (Martins et al., 2009),
however, no similar studies have been reported on marble goby.
In this study, marble goby was cultured in a fish rearing tank
treated by a RAS containing a hydroponic tank grown with leaf
vegetable (i.e. water spinach, Ipomoea aquatica). The influence of
the dimension ratio between the hydroponic tank and the fish
rearing tank (termed “component ratio”, i.e. hydroponic tank vol-
ume to rearing tank volume) on the fish growth, vegetable yield,
and nutrient removal was investigated; ammonia-N (TAN), nitrite-
N (NO2eN), nitrate-N (NO3eN), total-N (TN), 5-day biochemical
oxygen demand (BOD5), total suspended solid (TSS), and total
phosphorus (TP) were determined and assessed for this study. The
component ratio is an important process parameter to study as it
could have influence on the rate and extent of nutrient accumula-
tion and removal in the fish rearing tank, and also the nutrient
supplementation to the plants in the hydroponic tank; these two
factors could have effects on the production of both the fish and the
plants in the RAS. These evaluations are important to assess the
technical feasibility of using RAS as an alternative method for the
culture of the fish. So far, there has been little research reported on
the influence of the component ratio used in RAS, and existing
literature is limited to mostly studies performed on the use of RAS
for the culture of tilapia, catfish, tomato, and pak choi (Brassica
campestris L. subsp. chinensis) (Palm et al., 2014; Hu et al., 2015).
Materials and methods
Design of recirculating aquaponic system (RAS)
The RAS developed and used for this investigation is shown in
Fig. 1. It consists of fibreglass fish rearing tank, hydroponic tank,
denitrification sump, rapid sand filter tank coupled with filter floss
for solid removal, and aerated water reservoir tank.
Three fibreglass fish rearing tank were set up in series for the
culture of marble goby. Air stones connected to an air blower were
diffusely placed into the rearing tanks in order to maintain sufficient
oxygen supply to the fish. The water level in the rearing tanks was
maintained at a height of 0.85 m in order to achieve a water volume
of about 1000 L. The loss of water from evaporation, transpiration,
and sludge removal was replenished with water from the aerated
water reservoir tank. The rearing tanks were covered with 20 mm
plastic net to prevent the fish from jumping out of the tanks. The
temperature, level of dissolved oxygen (DO), and pH of water in the
rearing tanks were performed in situ through the use of YSI multi-
Please cite this article in press as: Lam, S.S., et al., Biological nutrient removal by recirculating aquaponic system: Optimization of the dimension
ratio between the hydroponic & rearing tank components, International Biodeterioration & Biodegradation (2015), http://dx.doi.org/10.1016/
j.ibiod.2015.03.012
probe meter (YSI 550A model) and pH meter (Cyber Scan water-
proof). The water temperature was found to be in the range of
27e28  C, and the pH was in the range of 7e8, and the level of DO
was monitored at >6 mg/L; the water quality parameters were re-
ported to be suitable for the culture of marble goby (Lam et al., 2008).
Water effluent from the fish rearing tanks was first treated by an
aerated hydroponic tanks (1e10 m2 area, 0.5 m height of water)
grown with leaf vegetable (i.e. water spinach, Ipomoea aquatic). The
effluent was sprinkled over the vegetables in the hydroponic tank
where the undesirable nutrient wastes (e.g. ammonia, nitrite, ni-
trate) produced from both the fish and the fish feed were absorbed
and recovered by the plant as nutrient. Water spinach were planted
on polystyrene sheets that floated along the hydroponic tank; the
polystyrene sheet supported the plants at the water surface with
roots suspended in the culture water, providing good exposure of
the roots to the culture water while preventing undesired clogging.
The effluent from the hydroponic tank was then allowed to
trickle down to a well-sealed sump for denitrification treatment.
The denitrification sump was incubated with denitrifying bacteria
in order to convert the remaining nitrate in the effluent to nitrogen
gas under anaerobic condition. The different components in the
RAS were assembled at different heights such that the water was
flowed under gravity. The effluent from the denitrification sump
was then pumped vertically to the rapid sand filter tank filled with
sands and filter floss of different thicknesses for solid removal.
Finally, the effluent from the sand filter tank was channelled into an
aerated water reservoir tank filled with crushed coral, which serves
as a buffering substrate to maintain a pH of 7.5 ± 0.5 in the RAS
system. The treated water was then returned to the fish rearing
tank by gravity. PVC pipelines were used to circulate water between
the fish rearing tank and the RAS components. The water flow rates
were controlled by the gate valves installed in the RAS.
Fish and acclimation
Marble goby with a body weight of approximately 100 g was
obtained from various local sources. Before experiments, the fishes
were acclimated individually in fibreglass holding tanks supplied
with a continuous flow of well-aerated fresh water (water tem-
perature: 27  C; pH: 7.5; DO > 6 mg/L). The water was passed
through a UV disinfection unit before being supplied to the holding
tanks in order to ensure the water is disease free. During acclima-
tion, the fish were fed to satiation once a day with live tilapia
(Oreochromis niloticus). Live tilapia was selected for this study due
to its ease of acquisition and its suitability as a feed to marble goby
as has been reported by previous studies (Lam et al., 2014), and the
fact that marble goby is a passive carnivorous fish and there is
virtually no suitable artificial feed for the fish.
Experimental details on the influence of component ratio
Experimental procedure
The experiment was conducted in fish rearing tanks treated by
RAS (see Section 2.1) with the water temperature maintained at
27  C, pH controlled at 7.5, and the level of DO monitored at > 6 mg/
L. The fish rearing tanks were subjected to natural 24-h light: dark
cycle (i.e. 12 h of light photoperiod: 12 h of dark photoperiod).
The variable to be studied was the dimension ratio between the
hydroponic tank and the fish rearing tank (termed “component
ratio”, i.e. hydroponic tank volume to rearing tank volume). Six
component ratios of hydroponic tank volume to rearing tank vol-
ume were selected and assessed for this study, namely ½ m3:1 m3,
1 m3:1 m3, 2 m3:1 m3, 3 m3:1 m3, 4 m3:1 m3, and 5 m3:1 m3
(Table 1). The different hydroponic tank volumes were achieved
through the use of 6 hydroponic tanks with different areas (i.e.
 S.S. Lam et al. / International Biodeterioration & Biodegradation xxx (2015) 1e9 3

Fig. 1. Schematic layout of recirculating aquaponic system. A: fish rearing tank; B: hydroponic tank planted with water spinach on polystyrene sheets; C: hydroponic tank with no
plants as control; D: water sprinkler, E: denitrification sump; F: pump; G: rapid sand filter tank with filter floss; H: water reservoir tank with crushed coral; I: air blower; J: Gate
valve.

Table 1
Selected component ratioa between the hydroponic and rearing tank.

Area planted with water Height of hydroponic tank Volume of hydroponic tank Volume of rearing tankb Component ratio
m2 m m3 m3 m3:m3

1 0.5 0.5 1 0.5:1

2 0.5 1 1 1:1
4 0.5 2 1 2:1
6 0.5 3 1 3:1
8 0.5 4 1 4:1
10 0.5 5 1 5:1
a
The dimension ratio between the hydroponic tank and the fish rearing tank, i.e. hydroponic tank volume to rearing tank volume.
b
The rearing tank has an area of 1.18 m2 and a water height of 0.85 m.

1 m2, 2 m2, 4 m2, 6 m2, 8 m2, 10 m2), respectively. All the hydroponic
tanks were filled with water to a height of 0.5 m in order to produce
the 6 desired hydroponic tank volumes (e.g. 1 m3 of hydroponic
tank volume was prepared by filing the 2 m2 hydroponic tank with
0.5 m height of water, i.e. 2 m2  0.5 m ¼ 1 m3). Polystyrene sheets
measured at 10 cm  10 cm were then distributed into the hy-
droponic tanks to allow floating along the hydroponic tank.
15 pieces of marble goby with an initial average body weight of
100 ± 5 g were randomly placed into three replicate fibreglass fish
rearing tanks, respectively. Prior to experimentation, the fishes
were acclimated to the rearing tank conditions and fed to satiation
once a day with live tilapia for 2 weeks; the selected feed was
analysed for crude protein, crude fat, ash, and fibre content ac-
cording to AOAC (1990) and these were presented in Table 2.
Seedlings of pre-weighed water spinach were planted directly into
the polystyrene sheets that floated along the hydroponic tank with
the desired hydroponic tank volume. Then, the experiments
commenced with the test of one component ratio at a time and
each component ratio was tested with the fishes cultured in rearing
tank for a period of 30 days. Each treatment of component ratio was
Table 2
Proximate composition (wt.%) of live tilapia.
Proximate composition as fed basisa
replicated thrice and also compared to the hydroponic tank with no
plants as a control.
During the 30-day experimental period, the fishes were fed once
a day to satiation (equivalent to a feeding rate of about 2e4% body
weight/day) with live tilapia. The feed consumption and fish mor-
tality were recorded daily. Water samples were collected at a daily
interval from the inlet and outlet of each rearing tank, the hydro-
ponic tank, the denitrification sump, the sand filter tank, and the
water reservoir tank. The level of DO, pH, and temperature of the
water samples were also recorded when the sampling was per-
formed. The water samples were analysed for ammonia-N, nitrite-
N, nitrate-N, total-N, 5-day biochemical oxygen demand, total sus-
pended solid, and total phosphorus for determination of the water
quality and the levels of nutrient wastes in the different RAS com-
ponents. These evaluations are important to assess the efficiency of
the RAS components in removing the nutrient wastes with a view to
assessing the overall performance of the RAS system. The sampling
was performed at between 8.30 am and 9.30 am daily and the water
samples were then transferred into glass bottles and stored at 4  C
until further analysis. At the end of the 30-day experimental period,
the fishes in the rearing tank and the water spinach were collected
and subjected to different growth and weight measurements. The
same procedure was repeated with different component ratios until
Live tilapia all the proposed component ratios were tested.

Moisture 76
Crude protein 16 Measurement of fish growth and vegetable yield
Crude fat 4 The growth and survival of fishes were monitored on placing the
Ash 4 fishes into the rearing tanks till the fishes were harvested at the end
a
Means of three biological replicates. of the experimental period. At the end of the experimental period,

Please cite this article in press as: Lam, S.S., et al., Biological nutrient removal by recirculating aquaponic system: Optimization of the dimension
ratio between the hydroponic & rearing tank components, International Biodeterioration & Biodegradation (2015), http://dx.doi.org/10.1016/
j.ibiod.2015.03.012
4 S.S. Lam et al. / International Biodeterioration & Biodegradation xxx (2015) 1e9
the fishes in the rearing tanks were individually weighed and
measured for both their growth and length in order to assess the
overall performance of the RAS system. Then, three fishes from
each rearing tank were randomly collected and frozen. The three
fishes were dried at 60  C, grounded, and analysed for proximate
composition according to AOAC (AOAC, 1990). Data on initial
weight, final weight, feed intake, and proximate composition of
diets were used to calculate the specific growth rate (SGR), absolute
growth rate (AGR), and feed conversion efficiency (FCE). These were
calculated as below:
%SGRð%=dayÞ ¼ ½ðlnW2  lnW1 Þ=ðT2  T1 Þ  100
AGR ¼ ðW2  W1 Þ=T2  T1
Where:
W1 ¼ Mean initial wet body weight of fish (g)
W2 ¼ Mean final wet body weight of fish (g)
T2  T1 ¼ Length of culture period, day (d)
Feed conversion ratio ðFCRÞ ¼ Total feed intakeðgÞ=
Weight gain of fishðgÞ
The growth of water spinach was monitored weekly by
measuring the height of the leaf vegetables and calculating the
number of the green shoots on the planted area in the hydroponic
tank. The water spinach was harvested when the vegetable had
achieved a height of about 50 cm. The water spinach was then
measured for their weight and the total weight of the water spinach
produced throughout the 30-day experimental period was
recorded.
Measurement of water quality and analysis of nutrient
concentration
The water samples were analysed for ammonia-N (TAN), nitrite-
N (NO2eN), nitrate-N (NO3eN), total-N (TN), 5-day biochemical
oxygen demand (BOD5), total suspended solid (TSS), and total
phosphorus (TP). Measurements of water temperature, pH, and DO
were performed in situ using the YSI multi-probe meter (YSI 550A
model) and pH meter (Cyber Scan Waterproof), respectively. TAN,
NO2eN, NO3eN, TN, and TP were analysed using standard method
adapted by Parsons et al. (Parsons et al., 1984), whereas TSS and
BOD5 concentrations were determined in accordance with the
standard methods (APHA, 1995). The water samples that could not
be analysed immediately were frozen at 15  C for a maximum of 1
week; this was within the recommended maximum storage time of
2 weeks by Parsons et al. (Parsons et al., 1984).
Statistical analyses
Data analyses were performed using the SPSS ver. 16.0 statistical
package (SPSS Inc. USA) and Microsoft Excel assuming statistical
significance if P < 0.05 (a set at 0.05). Data are calculated as
mean ± standard deviation (SD) and were subjected to one-way
analysis of variance (ANOVA) (Steel and Torrie, 1980). If signifi-
cant statistical differences were indicated at the 0.05 levels, then
Duncan's multiple range test was used to compare means in order
to identify significant differences between means (Duncan, 1955).
Results and discussion
Growth, survival, and production of marble goby
Fig. 2 and Fig. 3 show the growth, survival, and production of
marble goby under different component ratios. Component ratio
has a significant influence on the growth, survival, and production
Please cite this article in press as: Lam, S.S., et al., Biological nutrient removal by recirculating aquaponic system: Optimization of the dimension
ratio between the hydroponic & rearing tank components, International Biodeterioration & Biodegradation (2015), http://dx.doi.org/10.1016/
j.ibiod.2015.03.012
of the fish. The results show that higher growth was obtained at
higher component ratios as indicated by higher absolute and spe-
cific growth rate. The growth of fish observed at a component ratio
of
3 m3/m3 was significantly higher than those of fish cultured at
lower component ratios. In addition, the fish cultured at higher
component ratios (
3 m3/m3) demonstrated both higher produc-
tion and survival rate, recording a production of
70 g and a 100%
survival rate at the end of the 30-day experimental period. These
results are in agreement with the findings of McMurtry et al.
(1993a), McMurtry et al. (1993b) who reported that higher pro-
duction of fish was observed at increased ratio of the size of bio-
(1) filter tank grown with plants to the size of fish tank in a RAS sys-
tem culturing both tilapia and tomato.
(2) The higher component ratios, which contains a larger area
planted with more leaf vegetables (i.e. water spinach), provide
more plants to absorb the wastes generated from the culture of the
fish and in turn provide a good water quality condition to the fish.
The good water quality environment could prevent the occurrence
of undesired disease to the fish and this helps to explain the higher
growth and survival of fish observed at high component ratios. The
lowest component ratio that demonstrates the highest growth and
(3)
survival of the fish was observed at a component ratio of 3 m3/m3,
representing the optimum component ratio to obtain high pro-
duction of the fish from this RAS.
Feed intake and utilization by marble goby
The utilization of feed (i.e. live tilapia) by marble goby is pre-
sented in Table 3. It was observed that the amount of feed intake
increased with increasing component ratio. The higher feed intake
also corroborates the higher growth observed at higher component
ratios (Figs. 2 and 3), suggesting that the fish showed improved
eating behaviour and this had led to higher growth at higher
component ratios. It is thought that the better water quality con-
dition (Table 4 and Table 5) achieved at higher component ratios
ensures that the stress imposed on the marble goby was kept to a
minimum as the fish rearing tank was maintained under safe levels
of nutrient concentration and as a result the fish was not exposed to
toxic condition and less likely to be infected with disease. In
addition, the use of live tilapia as the feed to the marble goby was
likely to have improved the intake of feed by the fish. Live tilapia
has been reported as the preferential diet for marble goby due to
the high ingested protein quality of this diet for marble goby
nutrition, supplying the fish with proper amino acids for meta-
bolism and muscle growth (Laurel and Garling, 1997; Lam et al.,
2014).
However, there was no difference in the feed conversion ratio
(FCR) obtained with the different component ratios. This was likely
due to the use of same feed (i.e. live tilapia) in all the experiments,
and thus providing the similar nutrition and quality of protein for
the growth of fish. The results show a FCR of about 1.5, which shows
a better value compared to the FCR reported for marble goby fed
with soybean meal, which ranged from 1.8 to 4 (Yong et al., 2013),
probably due to the preference of marble goby as a carnivorous fish
for live food and also the better protein quality provided by the live
tilapia for growth.
Production of nutrient waste and water quality in fish rearing tank
The mean concentrations of nutrient wastes (i.e. TAN, NO2eN,
NO3eN, TN, TP) and water quality parameters (TSS, BOD5, DO, pH,
temperature) in the fish rearing tank were determined and the
results are presented in Tables 4 and 5. The highest concentrations
of nutrient wastes, TSS, and BOD5 were observed at the component
ratio of 0.5 m3/m3. This could be attributed to the comparatively
 S.S. Lam et al. / International Biodeterioration & Biodegradation xxx (2015) 1e9 5

Fig. 2. Growth of marble goby in relation to different component ratios. The data presented show typical results for the growth of marble goby obtained in experiments with low
standard deviations (i.e. a maximum error rate of ±5%) shown in the values for each reference point.

Fig. 3. Production and survival of marble goby in relation to different component ratios. The data presented show typical results for the production and survival of marble goby
obtained in experiments with low standard deviations (i.e. a maximum error rate of ±5%) shown in the values for each reference point.

Table 3
Feed intake (as fed) and utilization of marble goby.

Component ratio Total feed intake (g) Average feed intake (%BW/day) FCR

0.5 m3:1 m3 69 ± 4 a
2.3 ± 0.2 a
1.5 ± 1 a

1 m3:1 m3 80 ± 5 b
2.7 ± 0.2 b
1.5 ± 1 a

2 m3:1 m3 93 ± 3 c
3.1 ± 0.1 c
1.6 ± 1 a

3 m3:1 m3 108 ± 5 d
3.6 ± 0.2 d
1.5 ± 1 a

4 m3:1 m3 109 ± 4 d
3.6 ± 0.2 d
1.5 ± 1 a

5 m3:1 m3 111 ± 6 d
3.7 ± 0.3 d
1.5 ± 1 a

Values are mean from triplicate data (n ¼ 3) and means with the same superscript in the same column are not statistical different (P > 0.05).
Feed conversion ratio (FCR) ¼ Total feed intake (g)/Weight gain of fish (g).

Table 4
Mean concentrations of nutrient wastes in fish rearing tank.

Component ratio TAN (mg/L) NO2eN (mg/L) NO3eN (mg/L) TN (mg/L) TP (mg/L)

0.5 m3:1 m3 25 ± 2 a
0.8 ± 0.2 a
43 ± 2 a
70 ± 2 a
21 ± 1 a

1 m3:1 m3 22 ± 1 b
0.7 ± 0.1 a
39 ± 2 a
63 ± 2 b
18 ± 1 bc

2 m3:1 m3 19 ± 1 c
0.7 ± 0.1 a
34 ± 2 b
55 ± 3 c
16 ± 1 c

3 m3:1 m3 15 ± 2 d
0.5 ± 0.1 b
24 ± 2 c
40 ± 2 d
14 ± 1 d

4 m3:1 m3 14 ± 1 d
0.5 ± 0.1 b
20 ± 3 cd
37 ± 2 d
13 ± 2 d

5 m3:1 m3 14 ± 2 d
0.5 ± 0.1 b
19 ± 1 d
37 ± 2 d
12 ± 1 d

Values are mean from triplicate data (n ¼ 3) and means with the same superscript in the same column are not statistical different (P > 0.05).

Please cite this article in press as: Lam, S.S., et al., Biological nutrient removal by recirculating aquaponic system: Optimization of the dimension
ratio between the hydroponic & rearing tank components, International Biodeterioration & Biodegradation (2015), http://dx.doi.org/10.1016/
j.ibiod.2015.03.012
6 S.S. Lam et al. / International Biodeterioration & Biodegradation xxx (2015) 1e9

Table 5
Water quality parameters in fish rearing tank.

Component ratio TSS (mg/L) BOD5 (mg/L) DO (mg/L) pH (mg/L) Temperature (mg/L)

0.5 m3:1 m3 43 ± 2 a
5.9 ± 0.1 a
6.1 ± 0.1 a
6.5 ± 0.1 a
27.3 ± 0.3 a

1 m3:1 m3 35 ± 1 b
5.6 ± 0.1 b
6.5 ± 0.2 b
6.8 ± 0.1 b
27.9 ± 0.3 a

2 m3:1 m3 32 ± 1 c
5.4 ± 0.1 c
6.6 ± 0.1 b
7.2 ± 0.2 b
28.4 ± 0.4 b

3 m3:1 m3 28 ± 1 d
5.2 ± 0.1 d
6.9 ± 0.2 c
7.5 ± 0.1 c
27.6 ± 0.2 a

4 m3:1 m3 27 ± 2 d
5.2 ± 0.1 d
7.3 ± 0.1 c
7.6 ± 0.1 c
28.2 ± 0.3 b

5 m3:1 m3 27 ± 1 d
5.1 ± 0.1 d
7.2 ± 0.1 c
7.6 ± 0.1 c
27.4 ± 0.2 a

Values are mean from triplicate data (n ¼ 3) and means with the same superscript in the same column are not statistical different (P > 0.05).

poor capability of the RAS to treat and remove those generated
wastes and resulted in accumulation of those wastes in the fish
rearing tank; this is likely to account for the higher concentration of
those wastes observed at 0.5 m3/m3.
It was revealed that lower concentrations of nutrient wastes,
TSS, and BOD5 were obtained at higher component ratios (
3 m3/
m3), which once again corroborates the good water quality condi-
tions provided to the culture of the fish at higher component ratios.
However, there was no significant difference between the nutrient
wastes, TSS, and BOD5 obtained from the component ratio of 3 m3/
m3, 4 m3/m3, and 3 m3/m3, suggesting that a minimum component
ratio of is 3 m3/m3 is sufficient to maintain a similar water quality
environment to the culture of the fish.
The results on production of nutrient wastes correlate well with
the results of growth and feed utilization (Fig. 2 and Table 3). In this
study, higher growth and feed intake was obtained at higher
component ratios. This indicates that more nutrients were pro-
vided and utilised for the growth of the fish, and resulted in less
nutrients being excreted or discarded as undesirable nutrient
wastes. It is generally acceptable that the excretion of nutrient
wastes (e.g. TAN) generated from catabolism would be reduced as
nutrient retention (e.g. protein) is increased. This helps to explain

Fig. 4. Growth rate (A) and total yield (B) of water spinach. Growth rate ¼ height of vegetable growth/day. The data presented show typical results for the growth rate and total yield
of water spinach obtained in experiments with low standard deviations (i.e. a maximum error rate of ±5%) shown in the values for each reference point.

Please cite this article in press as: Lam, S.S., et al., Biological nutrient removal by recirculating aquaponic system: Optimization of the dimension
ratio between the hydroponic & rearing tank components, International Biodeterioration & Biodegradation (2015), http://dx.doi.org/10.1016/
j.ibiod.2015.03.012
 S.S. Lam et al. / International Biodeterioration & Biodegradation xxx (2015) 1e9 7

the lower concentrations of nutrient wastes observed at higher
component ratios. On the whole, it was shown in this study that
component ratio had a significant influence on the production of
nutrient wastes.
The BOD5 detected in this study ranged from 5.1 to 5.9 mg/L. The
BOD5 was likely to be derived from the fish metabolism and also the
increase of dissolved biodegradable organic substances and oxygen
demand associated with the decomposition of the fish feces and the
uneaten feed. Fish feces and uneaten feed were produced during
the experimental period, and these materials were likely to account
for the BOD5 detected in this study. The BOD5 detected has
exceeded the acceptable 5 mg/L of BOD5 limit for aquaculture by
Law (1987). This suggests that a bio-filtration or hydroponic tank
must be provided to the RAS for the removal of the dissolved
biodegradable organic substances in order to further reduce the
BOD5 in the water.
In this study, the TSS concentration determined in this study
ranged from 27 to 43 mg/L. The TSS was likely to be comprised of
the uneaten feed and the faeces excreted by the fish. The TSS
determined at the component ratio of 0.5 m3/m3 was significantly
higher than those determined at higher component ratio. It was
visually observed that higher amounts of uneaten feed were spitted

Fig. 5. Performance of the hydroponic tank of RAS in removing the nutrient wastes (TAN, NO2eN, NO3eN, TN, TP), TSS, and BOD5 under different component ratios (hydroponic
tank volume: rearing tank volume). Removal (%) by RAS ¼ (Before treatment e After treatment)*100%/Before treatment. The data presented show typical results for the perfor-
mance of the hydroponic tank of RAS obtained in experiments with low standard deviations (i.e. a maximum error rate of ±5%) shown in the values for each reference point.

Please cite this article in press as: Lam, S.S., et al., Biological nutrient removal by recirculating aquaponic system: Optimization of the dimension
ratio between the hydroponic & rearing tank components, International Biodeterioration & Biodegradation (2015), http://dx.doi.org/10.1016/
j.ibiod.2015.03.012
8 S.S. Lam et al. / International Biodeterioration & Biodegradation xxx (2015) 1e9
out from the mouth of marble goby cultured at this component
ratio, probably due to the low feed intake and the poor feeding
behaviour of the fish under a stressed and poor water quality
condition (see Table 3 and Section 3.2); this was likely to account
for the higher TSS.
In general, low TSS was observed in this study. However, the TSS
(27e43 mg/L) had exceeded the recommended limit of 15 mg/L for
warm water fishes in recirculating aquaculture systems, and also
the tentative upper limit of 25 mg/L for freshwater fish (Timmons
et al., 2002). A high TSS could damage the fish gills and harbour
pathogens that are detrimental to fish health. This implies a need to
include a solid removal mechanism in the RAS. A rapid sand filter
could contribute in reducing both the TSS and BOD5 to the safe
limits as previously mentioned.
In addition to TSS and BOD5, the water temperature was found
to be in the range of 27e28  C, and the pH was in the range of 7e8,
and the level of DO was monitored at
6 mg/L. These water quality
parameters were found to be within the suitable levels for the
culture of marble goby (Lam et al., 2008) and also within the ideal
levels for RAS (Timmons et al., 2002).
Vegetable yield and nutrient removal by RAS
Fig. 4 shows the growth rate and yield of water spinach under
different component ratios. The water spinach was observed to
have flourished over the experimental period and showed no signs
of mineral imbalances. At end of the experimental period, the
vegetables recorded a height of about 50 cm. The growth and the
yield of the vegetable were found to increase with increasing
component ratio, but the growth and the yield showed a decrease
after a component ratio of 3 m3/m3, suggesting that nutrient de-
ficiencies might have occurred due to insufficient nutrient wastes
available for the comparatively higher amounts of water spinach
planted in the hydroponic tank at 4 and 5 m3/m3, resulting in a
decline in nutritional supplementation to the vegetables and the
subsequent growth and yield of the vegetables. The highest vege-
table yield (i.e. 22 kg/harvest) was observed at a component ratio of
3 m3/m3, suggesting that this represents the optimum balance
between sufficiently high nutrient supplementation to vegetable
yield, while not being so low that the RAS was unable to remove the
nutrients and resulted in rapid nutrient accumulation and tox-
ification in the fish rearing tank (See Fig. 5 for the removal effi-
ciency of nutrient wastes by RAS).
Fig. 5 demonstrates the performance of the hydroponic tank of
RAS in removing the nutrient wastes (TAN, NO2eN, NO3eN, TN, TP),
TSS, and BOD5 under different component ratios. Component ratio
was found to have a significant influence on the overall nutrient
removal. The removal of nutrient wastes, TSS, and BOD5 was found
to increase with increasing component ratio. In particular, high
removal efficiency (>70%) was achieved for TAN, NO2eN, NO3eN,
TN, and TSS at high component ratios (
3 m3/m3), except for TP and
BOD5 that record a removal efficiency of 60% and 63%, respectively.
The removal of nutrient wastes, TSS, and BOD5 could occur via
several mechanisms, e.g. the absorption of nutrient wastes and the
retention of TSS by vegetable planted in the hydroponic tank
through the root system, the removal of dissolved solids via mi-
crobial assimilation by microorganisms in the water column, or the
retention and adsorption on bio-films formed within the root sys-
tem of vegetable planted in the hydroponic tank (Timmons et al.,
2002). The results demonstrate that the water treated by the RAS
at a component ratio of
3 m3/m3 showed an acceptable water
quality with low levels of nutrient wastes (i.e. 3 mg/L of TAN,
0.1 mg/L of NO2eN, 7 mg/L of NO3eN), TSS (i.e. 3 mg/L), and
BOD5 (i.e. 2.4 mg/L); the recommended limit for TAN, NO2eN,
NO3eN, TSS, and BOD5 is 3 mg/L, 1 mg/L, 100 mg/L, 15 mg/L, and
Please cite this article in press as: Lam, S.S., et al., Biological nutrient removal by recirculating aquaponic system: Optimization of the dimension
ratio between the hydroponic & rearing tank components, International Biodeterioration & Biodegradation (2015), http://dx.doi.org/10.1016/
j.ibiod.2015.03.012
5 mg/L, respectively (Law, 1987; Timmons et al., 2002). Although
the current set up of the RAS provides relatively safe levels of
nutrient wastes, were the concentrations assessed to have
increased and pose too great a risk, or if reduction of the nutrient
wastes to lower levels was required, effort could be made to reduce
the number of fish cultured in the rearing tank, or the polystyrene
sheets that floated along the hydroponic tank could be planted with
more water spinach through reduction of the area of spacing for the
plants (e.g. from 10 cm  10 cm spacing to 8 cm  8 cm spacing).
In this study, higher fish growth, feed intake, vegetable yield,
and nutrient removal were observed at a component ratio of
3e5 m3/m3. However, there was no significant difference between
the removal of nutrient wastes, TSS, and BOD5 obtained from those
component ratios. This suggests that a minimum component ratio
of is 3 m3/m3 is adequate to provide the similar results for the
culture of the fish in RAS. Combined with the results showing the
highest growth and yield of water spinach, this suggests that 3 m3/
m3 represents the optimum component ratio to obtain high pro-
duction of both the fish and the vegetable from the RAS.
Conclusions
Component ratio was found to have a significant influence on
nutrient removal and production of marble goby and water spinach
in RAS. A component ratio of
3 m3 of hydroponic tank volume to
1 m3 of fish rearing tank volume showed advantages in improving
the production of the fish and vegetable and removing the nutrient
wastes, TSS, and BOD5 generated from the culture of the fish. The
results indicate that RAS show exceptional promise as a means to
the reduction of biological nutrients accumulated in aquaculture
wastewater and in turn providing a good water quality environ-
ment for fish culture.
Acknowledgements
Su Shiung Lam acknowledges the technical and financial assis-
tance by Ministry of Science, Technology, and Innovation of
Malaysia Government, Institute of Tropical Aquaculture, and Uni-
versity Malaysia Terengganu.
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